The pathogenesis of endometriosis and adenomyosis: insights from single-cell RNA sequencing†.

Endometriosis and adenomyosis are two similar gynecological diseases that are characterized by ectopic implantation and the growth of the endometrial tissue. Previous studies have reported that they share a common pathophysiology in some respects, such as a similar cellular composition and resistance to the progestogen of lesions, but their underlying mechanisms remain elusive. Emerging single-cell ribonucleic acid sequencing (scRNA-seq) technologies allow for the dissection of single-cell transcriptome mapping to reveal the etiology of diseases at the level of the individual cell. In this review, we summarized the published findings in research on scRNA-seq regarding the cellular components and molecular profiles of diverse lesions. They show that epithelial cell clusters may be the vital progenitors of endometriosis and adenomyosis. Subclusters of stromal cells, such as endometrial mesenchymal stem cells and fibroblasts, are also involved in the occurrence of endometriosis and adenomyosis, respectively. Moreover, CD8+ T cells, natural killer cells, and macrophages exhibit a deficiency in clearing the ectopic endometrial cells in the immune microenvironment of endometriosis. It seems that the immune responses are activated in adenomyosis. Understanding the immune characteristics of adenomyosis still needs further exploration. Finally, we discuss the application of findings from scRNA-seq for clinical diagnosis and treatment. This review provides fresh insights into the pathogenesis of endometriosis and adenomyosis as well as the therapeutic targets at the cellular level.

Preoperative Prevalence and Risk Factors For Calf Muscular Vein Thrombosis in Elderly Patients with Hip Fracture.

OBJECTIVE: Increasing evidence has shown that calf muscular vein thrombosis (CMVT) can develop into proximal deep vein thrombosis, even causing pulmonary embolism. However, opinions about the prevalence and risk factors are still controversial. This study aimed to investigate the prevalence and risk factors for CMVT in elderly patients with hip fractures to facilitate their preoperative management. METHODS: We included 419 elderly patients with hip fracture who were treated in the orthopaedic department of our hospital from June 2017 to December 2020. The patients were divided into CMVT and non-CMVT groups based on color Doppler ultrasound screening of the venous system in the lower extremities. Clinical data, such as age, sex, body mass index, time from injury to admission, and laboratory data were collected. Univariate and multivariate logistic regression analyses were performed to determine independent risk factors for CMVT. A receiver operating characteristic curve was used to analyze the predictive effectiveness of the model. Finally, the clinical utility of the model was analyzed using decision curve analysis and clinical impact curves. RESULTS: The prevalence of preoperative CMVT was 30.5% (128/419). Independent predictors of preoperative CMVT identified by univariate and multivariate logistic regression analyses were sex, time from injury to admission, American Society of Anesthesiologists (ASA) classification, C-reactive protein (CRP) level, and D-dimer level (p < 0.05). The area under curve (AUC) was 0.750 (95% CI: 0.699-0.800, p < 0.001) and the sensitivity and specificity were 0.698 and 0.711, respectively, which meant the prediction model has a good efficacy in the prediction of CMVT risk. In addition, the fitting degree of the prediction model was also good (Hosmer-Lemeshow χ2 = 8.447, p > 0.05). The clinical utility of the model was verified using decision curve analysis and clinical impact curves. CONCLUSION: Sex, time from injury to admission, ASA classification, CRP level, and D-dimer levels are independent preoperative predictors of CMVT in elderly patients with hip fractures. Measures should be taken for patients with these risk factors to prevent the occurrence and deterioration of CMVT.

The results of ureteral stenting after ureteroscopic lithotripsy for ureteral calculi: a systematic review and meta-analysis.

PURPOSE: We evaluated the necessity and adverse effects of routine ureteral stent placement after ureteroscopic lithotripsy for ureteral stones. MATERIALS AND METHODS: A systematic search of PubMed®, Embase® and the Cochrane Library was performed to identify all randomized controlled trials. All relevant studies were on the outcomes and complications of ureteroscopic lithotripsy in the management of ureteral stones with or without a Double-J stent. The outcomes and complications included stone-free rate, operative time, lower urinary tract symptoms, hematuria, fever, urinary tract infection, pain and analgesia, unplanned medical visits and late postoperative complications. The Cochrane Collaboration Review Manager software (RevMan 5.0.2) was used for statistical analysis. RESULTS: A total of 16 randomized controlled trials were enrolled for analysis and involved 1,573 patients. Of these patients 797 were in the nonstented group and 776 in the stented group. There was a statistically significant difference in mean operative time between the 2 groups. The incidence of lower urinary tract symptoms and pain was significantly higher in the stented group than in the nonstented group. Significant differences between the groups were not found in fever, urinary tract infection, need for analgesia, unplanned readmission and late postoperative complications. CONCLUSIONS: This systematic review reveals the obvious disadvantages of ureteral stents after ureteroscopic lithotripsy in lower urinary tract symptoms and pain. Stents do not improve stone-free rate, fever, incidence of urinary tract infection, unplanned medical visits, requirement for analgesia and late postoperative complications. Ureteral stenting after uncomplicated ureteroscopic lithotripsy could be unnecessary.

Differential regulation of P2X(3) mRNA expression by peripheral nerve injury in intact and injured neurons in the rat sensory ganglia.

The P2X(3) receptor is a ligand-gated cation channel activated by the binding of extracellular adenosine 5'-triphosphate (ATP), an agent that has been suggested to have a role in the nociceptive pathway after tissue and nerve injury. After peripheral nerve injury, both down regulation and up regulation of the P2X(3) receptor in sensory ganglion neurons have been observed. The purpose of this study was to examine the precise regulation of P2X(3) mRNA expression in primary sensory neurons after nerve injury. We used two nerve injury models in the rat, the transection of the tibial and common peroneal nerves and the transection of the infraorbital nerve, and observed dorsal root ganglion (DRG) and trigeminal ganglion neurons, respectively. P2X(3) mRNA in both neuron populations was detected by in situ hybridization with an oligonucleotide probe that was confirmed by Northern blot analysis. To identify axotomized neurons, we examined the expression of activating transcription factor 3 (ATF3), which is regarded as a neuronal-injury marker, using immunohistochemistry. In the DRG, the mean percentage of P2X(3) mRNA-labeled neurons relative to the total number of neurons increased from 32.7% in the naive rats to 42.7% at 3 days after injury. The mean percentage of P2X(3) mRNA-labeled neurons in ATF3 immunoreactive (ir) neurons was 29.5% at 3 postoperative days, which gradually decreased to 11.2% at 28 days after injury. In the trigeminal ganglion, the mean percentage of P2X(3) mRNA-labeled neurons was 36.9% at 3 days after injury, versus 26.0% in the naive rats. In the ATF3-ir neurons, the mean percentage of P2X(3) mRNA-labeled neurons was 25.3% at 1 postoperative day and was reduced to 6.1% at 28 postoperative days. The finding that P2X(3) mRNA in ATF3-ir neurons decreased significantly after injury indicates that axotomized neurons decreased the expression of P2X(3) mRNA, despite the increase in P2X(3) mRNA relative to the total number of sensory ganglion neurons. These data strongly suggest that P2X(3) mRNA expression increases in intact neurons and that P2X(3) mRNA in intact neurons may play a role in the pathomechanism of post-nerve injury in primary sensory neurons.

Contribution of injured and uninjured dorsal root ganglion neurons to pain behavior and the changes in gene expression following chronic constriction injury of the sciatic nerve in rats.

Neuropathic pain models, such as the chronic constriction injury (CCI) model, are partial nerve injury models where there exist both intact and injured peripheral axons. Recent studies suggested that dorsal root ganglion (DRG) neurons with intact axons also show the alteration of excitability and gene expression and might have some role in the pathophysiological mechanisms of neuropathic pain. The incidence of pain-related behavior after the CCI is unstable and variable. In the present study, we used activating transcription factor 3 (ATF3) expression as a neuronal injury marker, and analyzed a relationship between the number of axotomized neurons and the incidence of pain-related behavior. We divided all rats into three groups according to the percentage of ATF3-immunoreactive (IR) neurons, group 1 (<12.5%), group 2 (12.5-25%), and group 3 (>25%). We found that rats in groups 2 and 3 showed thermal hyperalgesia, whereas only the rats in group 2 developed tactile allodynia from the third day to the fourteenth day after surgery. Rats in group 1 did not show thermal hyperalgesia or tactile allodynia. The DRG neurons in group 2 contained ATF3-IR neurons mainly in medium- and large-sized neurons. In order to investigate brain-derived neurotrophic factor (BDNF) and gamma-aminobutyric acid(A)-receptor (GABA(A)-R) regulation in both intact and injured primary afferent neurons after the CCI, we used a double-labeling method with immunohistochemistry and in situ hybridization, as well as double immunofluorescent staining. The CCI induced an increased number of BDNF-labeled neurons in the ipsilateral DRG and the increase in BDNF expression was observed mainly in small- and medium-sized neurons that were mainly ATF3-negative. On the other hand, the number of GABA(A)-Rgamma2 subunit mRNA-positive neurons decreased in the ipsilateral DRG and GABA(A)-R- and ATF3-labeled neurons rarely overlapped. These changes in molecular phenotype in intact and injured primary afferents may be involved in the pathophysiological mechanisms of neuropathic pain produced by partial nerve injury.

Expression of neurotrophic factors in the dorsal root ganglion in a rat model of lumbar disc herniation.

A variety of molecules released by inflammatory reactions in the dorsal root and dorsal root ganglion (DRG) may play important roles in the pathology of neuronal abnormalities in lumbar disc herniation. In order to elucidate the pathophysiological mechanisms of painful radiculopathy, secondary to lumbar disc herniation, we evaluated pain-related behavior and the change of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) expression in the DRG and dorsal root using a rat model of lumbar disc herniation. In the nucleus pulposus (NP) group, the left L4/5 nerve roots were exposed after hemilaminectomies and autologous intervertebral discs, which were obtained from coccygeal intervertebral discs, were implanted on each of the exposed nerve roots without mechanical compression. Rats in the NP group, but not the sham-operated rats, developed mechanical allodynia on the ipsilateral hind paw for 1 day after surgery and showed a significant increase in the number of NGF-immunoreactive (IR) cells in the nerve root and DRG. NGF-IR cells in the nerve root and DRG included macrophages and Schwann cells, because these cells were labeled for NGF and ED-1 or glial fibrillary acid protein by dual immunostaining. A significant increase in the percentage of BDNF-IR neurons in the DRG was observed in the NP group at 3 days after surgery and the increase in BDNF mRNA expression was confirmed using in situ hybridization histochemistry and reverse transcription-polymerase chain reaction. We also injected NGF into the endoneurial space of the normal rat spinal nerve root and found that the NGF injection produced dose-dependent mechanical allodynia on the ipsilateral hind paw at 1 day after surgery and an increase in the percentage of BDNF-IR neurons in the DRG at 3 days after surgery compared to the group receiving saline injection. These findings suggest that in the lumbar disc herniation model, i.e. neuritis of the nerve root, increased NGF produced by the inflammatory responses in the dorsal root and DRG tissues may affect the production of BDNF in the DRG and may play important roles in the modulation of the dorsal horn neurons. These changes in neurotrophic factors in the primary afferents may be involved in the pathophysiological mechanisms of neuropathic pain produced by lumbar disc herniation.

Tracing the genetic history of the Chinese people: mitochondrial DNA analysis of aneolithic population from the Lajia site.

Ancient DNA analysis was conducted on the dental remains of specimens from the Lajia site, dating back 3,800-4,000 years. The Lajia site is located in Minhe county, Qinghai province, in northwestern China. Archaeological studies link Lajia to the late period of the Qijia culture, one of the most important Neolithic civilizations of the upper Yellow River region, the cradle of Chinese civilization. Excavations at the site revealed that the inhabitants died in their houses as the result of a sudden flood. The Lajia site provides a rare chance to study the putative families, all of whom died at the same instant. Possible maternal familial relationships were investigated through mitochondrial DNA (mtDNA) sequence analysis. Twelve sequences from individuals found in one house were assigned to only five haplotypes, consistent with a possible close kinship. Results from analyses of RFLP typing and HVI motifs suggest that the Lajia people belonged to the haplogroups B, C, D, M*, and M10. This study, combined with archaeological and anthropological investigations, provides a better understanding of the genetic history of the Chinese people.